Human defensin, alpha 1 (DEFA1) ELISA kit
Product Details
| Description |
The Human defensin, alpha 1 (DEFA1) ELISA kit enables quantification of human DEFA1 in solution, e.g. in serum, plasma, or urine. The product is more flexible to set up the ELISA assay. The assay is based on the Sandwich ELISA mechanism and enzyme-substrate chromogenic reaction. The intensity of the color is proportional to the DEFA1 content in the sample. This kit has been verified with nanogram-level of sensitivity, high specificity, precision, good linearity, high recovery, and lot-to-lot consistency. More validation details are described in the product instructions.
DEFA1, also called HNP-1, is one of the four types of defensins found in the azurophilic granules of polymorphonuclear neutrophils. It is likely to play a role in phagocyte-mediated host defense. Studies have demonstrated that DEFA1 exhibits potent in vitro bactericidal activity against a broad range of microbes, including mycobacterial species. It is also involved in the endothelial cell dysfunction at the time of early atherosclerotic development. DEFA1 is usually up-regulated in inflammation, including sepsis and acute coronary.
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| Target Name |
defensin, alpha 1 |
| Alternative Names |
alpha 1 ELISA Kit; DEF1 ELISA Kit; DEF1_HUMAN ELISA Kit; DEFA1 ELISA Kit; DEFA1B ELISA Kit; DEFA2 ELISA Kit; Defensin 1 ELISA Kit; Defensin ELISA Kit; Defensin; alpha 1 ELISA Kit; Defensin; alpha 1; myeloid related sequence ELISA Kit; Defensin; alpha 2 ELISA Kit; HNP-1 ELISA Kit; HNP-2 ELISA Kit; HNP1 ELISA Kit; HP-1 ELISA Kit; HP-2 ELISA Kit; HP1 ELISA Kit; HP2 ELISA Kit; MRS ELISA Kit; Myeloid related sequence ELISA Kit; Neutrophil defensin 1 ELISA Kit; Neutrophil defensin 2 ELISA Kit |
| Abbreviation |
DEFA1 |
| Species |
Homo sapiens (Human) |
| Sample Types |
serum, plasma, urine |
| Detection Range |
0.625 ng/mL-40 ng/mL |
| Sensitivity |
0.156 ng/mL |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Immunology |
| Assay Principle |
quantitative |
| Measurement |
Sandwich |
| Precision |
| Intra-assay Precision (Precision within an assay): CV% |
| Three samples of known concentration were tested twenty times on one plate to assess. |
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| Inter-assay Precision (Precision between assays): CV% |
| Three samples of known concentration were tested in twenty assays to assess. |
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| Linearity |
| To assess the linearity of the assay, samples were spiked with high concentrations of human DEFA1 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. |
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Sample |
Serum(n=4) |
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| 1:1 |
Average % |
89 |
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| Range % |
83-94 |
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| 1:2 |
Average % |
84 |
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| Range % |
80-88 |
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| 1:4 |
Average % |
93 |
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| Range % |
89-97 |
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| 1:8 |
Average % |
95 |
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| Range % |
90-99 |
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| Recovery |
| The recovery of human DEFA1 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section. |
| Sample Type |
Average % Recovery |
Range |
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| Serum (n=5) |
97 |
94-102 |
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| EDTA plasma (n=4) |
90 |
86-96 |
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| Typical Data |
| These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
| ng/ml |
OD1 |
OD2 |
Average |
Corrected |
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| 40 |
2.058 |
2.014 |
2.036 |
1.933 |
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| 20 |
1.547 |
1.525 |
1.536 |
1.433 |
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| 10 |
1.104 |
1.071 |
1.088 |
0.985 |
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| 5 |
0.673 |
0.715 |
0.694 |
0.591 |
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| 2.5 |
0.459 |
0.437 |
0.448 |
0.345 |
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| 1.25 |
0.309 |
0.312 |
0.311 |
0.208 |
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| 0.625 |
0.205 |
0.194 |
0.200 |
0.097 |
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| 0 |
0.107 |
0.099 |
0.103 |
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| Materials provided |
- A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human DEFA1 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled DEFA1antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
- An instruction manual
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| Materials not provided |
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°C.
- Centrifuge
- Vortex
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Timer
- Test tubes for dilution
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| Storage |
Store at 2-8°C. Please refer to protocol. |
| Lead Time |
3-5 working days |
